control human dermal fibroblast cell line  (ATCC)

Journal: Antioxidants

Article Title: Bioenergetic Signatures of DLD Deficiency: Dissecting PDHc- and α-KGDHc-Linked Defects

doi: 10.3390/antiox15010019

Figure Lengend Snippet: Mitochondrial respiration is impaired in fibroblasts derived from patients with DLD deficiency. Mitochondrial oxygen consumption was assessed in controls (Ctrl1 and Ctrl2) and patient (Pt1–Pt6) fibroblasts using high-resolution respirometry (Oroboros O2k). ( A ) Routine respiration; ( B ) maximal respiration calculated as the difference between FCCP-stimulated and α-chaconine–permeabilized rates; ( C ) complex I-linked respiration (NADH-linked respiration, N-pathway), calculated as the difference between ADP and α-chaconine; ( D ) complex II-linked respiration (NS-pathway) calculated as the difference between respiration after rotenone and α-chaconine addition; ( E ) effect of complex I inhibition, calculated as the difference between FCCP-stimulated and rotenone-inhibited respiration; and ( F ) complex I/complex II respiration ratio (complex I-linked activity divided by complex II-linked activity). Each open circle represents an independent experimental run (N = 4–8 repeats per sample). All data were normalized to cell number. Statistical analysis was performed using the Mann–Whitney U test. * p < 0.05 vs. Ctrl1; numerical p values (0.05 < p < 0.1) are indicated on the plots. Abbreviations: Ctrl, control; Pt, patient.

Article Snippet: Dermal fibroblast primary cell lines from six genetically confirmed patients with DLD deficiency were obtained from the Pediatric Metabolic Disease Unit, Sheba Medical Center (IRB# SMC-21-8644, Figure 1, Table 1, and ), as well as two control cell lines: a control human dermal fibroblast cell line was purchased from ATCC (PCS-201-012; Ctrl 1, Manassas, VA, USA), and a primary cell line from a 39-year-old healthy male (Ctrl 2).

Techniques: Derivative Assay, Inhibition, Activity Assay, MANN-WHITNEY, Control

Journal: JBMR Plus

Article Title: Identification of cellular retinoic acid binding protein 2 (CRABP2) as downstream target of nuclear factor I/X (NFIX): implications for skeletal dysplasia syndromes

doi: 10.1093/jbmrpl/ziae060

Figure Lengend Snippet: QRT-PCR analysis of genes identified with altered expression in MEFs and human fibroblasts. QRT-PCR analysis in: Nfix +/+ , Nfix +/Del2 , Nfix Del2/Del2 , Nfix +/Del24 , Nfix Del24/Del24 , Nfix +/Del140 and Nfix Del140/Del140 MEFs; and 5 MSS (Del1, Del2, Del3, InsT and DupG) and 3 unrelated control (N1, N2 and N3) fibroblast cell lines, of (A) Crabp2 , (B) CRABP2, (C) Vcam1 , (D) VCAM1 , (E) Kctd12 , (F) KCTD12 , (G) Idi1 and (H) IDI1 expression, with Gapdh or GAPDH and Canx or CANX used as the housekeeping genes against which candidate gene expression was normalised. Data are represented as mean ± SEM, n = 4–12, * P <0 .05, * * P <0 .01, * * * * P <0 .0001.

Article Snippet: Human fibroblast cells were obtained from 5 MSS patients, as previously reported., In addition, 3 unrelated control human fibroblast cell lines (CRL2072, CRL2106, CRL1475) were obtained from ATCC (LGC Standards, Middlesex, UK).

Techniques: Quantitative RT-PCR, Expressing, Control, Gene Expression

Journal: JBMR Plus

Article Title: Identification of cellular retinoic acid binding protein 2 (CRABP2) as downstream target of nuclear factor I/X (NFIX): implications for skeletal dysplasia syndromes

doi: 10.1093/jbmrpl/ziae060

Figure Lengend Snippet: Western blot and densitometry analysis of genes identified with altered expression in MEFs and human fibroblasts. Western blot analysis in: (A) Nfix +/+ , Nfix +/Del2 , Nfix Del2/Del2 , Nfix +/Del24 , Nfix Del24/Del24 , Nfix +/Del140 and Nfix Del140/Del140 MEFs; and (B) 5 MSS (Del1, Del2, Del3, InsT and DupG) and 3 unrelated control (N1, N2, and N3) fibroblast cell lines. Quantified expression, using densitometry analysis, of (C-D) CRABP2, (E-F) VCAM1, (G-H) KCTD12 and (I-J) IDI1 expression. GAPDH and CANX were used as loading controls. Data are represented as mean ± SEM, n = 4, * P <0 .05, * * P <0 .01, * * * P <0 .001, * * * * P <0 .0001.

Article Snippet: Human fibroblast cells were obtained from 5 MSS patients, as previously reported., In addition, 3 unrelated control human fibroblast cell lines (CRL2072, CRL2106, CRL1475) were obtained from ATCC (LGC Standards, Middlesex, UK).

Techniques: Western Blot, Expressing, Control